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miR-365-3p Regulates Gastrointestinal Dysfunction in Diabetes Mellitus Rats via the TLR4/MyD88/NF-κB Pathway
Jiao Xiao1, Bin Gao1, Yan Xiao2
1Department of Endocrinology, Affiliated Nanhua Hospital, University of South China, Hengyang, China.
Abstract:
Over half of diabetes mellitus (DM) patients suffer from gastrointestinal motility disorders. miR-365-3p is involved in DM progression, but its role in gastrointestinal motility disorders remains unclear. This study explored whether miR-365-3p affects gastrointestinal motility in diabetic rats via the TLR4/MyD88/NF-κB pathway. A DM rat model was established using a high-fat, high-sugar diet and injected with a miR-365-3p mimic/inhibitor. DM symptoms, gastric emptying, intestinal propulsion rates, and gastrointestinal transit time were assessed. HE and TUNEL staining evaluated gastrointestinal pathology and apoptosis. qRT-PCR detected miR-365-3p levels, while ELISA assessed gastrointestinal motility-related factors. Immunofluorescence and Western blot analyzed C-kit, TLR4, and pathway proteins. DM rats exhibited increased body weight, blood glucose, and glucose intolerance, with reduced fasting insulin, confirming successful modeling. miR-365-3p was downregulated in DM rats. Injection of miR-365-3p mimic alleviated DM symptoms, reduced gastrointestinal tissue damage and apoptosis, and improved motility. The TLR4 agonist CRX-527 impaired these effects. In conclusion, miR-365-3p overexpression alleviates DM symptoms, gastrointestinal injury, and motility disorders by inhibiting the TLR4/MyD88/NF-κB pathway, offering a potential therapeutic target.
Insights
MicroRNA-365-3p (miR-365-3p) helps alleviate gastrointestinal issues in diabetes mellitus (DM) by inhibiting the TLR4/MyD88/NF-κB pathway. Restoring miR-365-3p levels improves gut motility and reduces damage in diabetic rats.
Area of Science:
- Endocrinology
- Gastroenterology
- Molecular Biology
Background:
- Gastrointestinal motility disorders affect over half of diabetes mellitus (DM) patients.
- The role of microRNA-365-3p (miR-365-3p) in DM-associated gastrointestinal dysfunction is not well understood.
Purpose of the Study:
- To investigate the effect of miR-365-3p on gastrointestinal motility in a rat model of diabetes mellitus.
- To elucidate the underlying molecular mechanism involving the TLR4/MyD88/NF-κB pathway.
Main Methods:
- Established a DM rat model using a high-fat, high-sugar diet.
- Administered miR-365-3p mimic or inhibitor to rats.
- Assessed gastric emptying, intestinal propulsion, and transit time.
- Utilized HE and TUNEL staining for histopathology and apoptosis analysis.
- Quantified miR-365-3p expression, motility factors, and pathway protein levels via qRT-PCR, ELISA, immunofluorescence, and Western blot.
Main Results:
- DM rats showed significantly altered metabolic parameters and impaired gastrointestinal motility.
- miR-365-3p was downregulated in DM rats.
- miR-365-3p mimic injection improved DM symptoms, reduced gastrointestinal injury and apoptosis, and enhanced motility.
- The TLR4 agonist CRX-527 reversed the beneficial effects of miR-365-3p mimic.
Conclusions:
- Overexpression of miR-365-3p ameliorates DM-induced gastrointestinal injury and motility disorders.
- miR-365-3p exerts its protective effects by inhibiting the TLR4/MyD88/NF-κB signaling pathway.
- miR-365-3p represents a potential therapeutic target for managing gastrointestinal complications in diabetes mellitus.
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